The DENV transcript is highlighted in red. ( B) Ribosome Profiling (Ribo-seq) and RNA-seq were performed in DENV-infected WT and EMC4-KO HEK293FT cells 18 hr post-infection to measure changes in translation efficiency (TE). The Ribosome profiling experiments were performed once for each condition. The red line represents fold-change of TE = 1.

Fold-change in TE for a given transcript is defined as the change in Ribosome Profiling RPKMs normalized to the change in RNA-seq RPKMs. Log 2 fold-change (FC) of reads per kb of transcript, per million mapped reads (RPKMs) between EMC4-KO and WT cells for Ribosome Profiling and RNA-seq are displayed on the y- and x-axis, respectively. RNA was treated with RNaseI and ribosome-protected footprints were purified, reverse transcribed and amplified for next-generation sequencing. ( A) Ribosome Profiling (Ribo-seq) and RNA-seq were performed in uninfected WT and EMC4-KO HEK293FT cells to measure changes in translation efficiency (TE). Cycloheximide was added to stop translation and total RNA was isolated. t-tests were performed to determine statistical significance and p-values are defined as ns = non significant, *<0.05, **<0.01, ***<0.001. In all figures, values are shown as mean of three biological replicates with standard deviation in (A)-(C), and as mean with standard error of the mean in (D). Viral RNA levels were normalized to 18S levels and data is displayed relative to the respective WT condition. Cells were infected with an moi of 0.5 for all viruses and harvested at 30hpi for ZIKV, 48hpi for DENV and WNV, and 72hpi for HCV. ( D) Quantitative RT-PCR of DENV, WNV, ZIKV and HCV RNA in WT or EMC4-KO Huh7.5.1 cells. Cells were infected with DENV-Luc, harvested at 48hpi and luminescence was measured. ( C) Replication of DENV in WT, EMC2- and EMC4-KO, and cDNA complemented KO HEK293FT cells. Dotted line indicates background from uninfected control cells.

( B) DENV infection of WT and EMC subunit KO HEK293FT cells. ( A) DENV infection of WT and EMC subunit KO Huh7.5.1 cells. Cells were infected with DENV-Luc, harvested at 30hpi and luminescence was measured.

How to change dashed line in flowjo 10